Robust Bioprocess Design and Evaluation of Commercial Media for the Serial Expansion of Human Induced Pluripotent Stem Cell Aggregate Cultures in Vertical-Wheel Bioreactors
This publication evaluates the robustness of a baseline process for serial expansion of human induced pluripotent stem cell aggregates in Vertical-Wheel bioreactors. Five commercial media and multiple cell lines were examined through short-term culture and serial passaging. Process outputs included growth kinetics, aggregate morphology, harvest efficiency, pluripotency, genomic stability, and response to stresses associated with in-vessel dissociation. All tested media supported spherical aggregate formation and strong expansion during an initial six-day culture, but important performance differences emerged only during serial passage. Some formulations maintained growth and cell quality over multiple cycles, including ten consecutive passages, while others produced cell lysis, reduced growth, or loss of aggregate morphology. The study demonstrates that robust process design requires long-term evaluation of media and operating variables rather than reliance on a single short culture.
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This publication reports a fully suspended process for expansion and six-stage differentiation of human induced pluripotent stem cells into stem cell-derived islets in 0.1 L and 0.5 L Vertical-Wheel® Bioreactors. A fivefold increase in vessel volume produced a twelvefold increase in islet-equivalent yield without compromising islet structure. The 27-day process generated endocrine populations expressing ?-, ?-, and ?-cell markers with glucose-responsive insulin secretion and transcriptomic characteristics approaching adult islets. The study also evaluated donor-derived iPSC lines, cellular…
This protocol provides step-by-step guidance for forming and expanding pluripotent stem cell aggregates in PBS-MINI Vertical-Wheel® bioreactors. The workflow begins with thawing and adherent seed-train culture in T-flasks, followed by transfer to three-dimensional suspension culture in 0.1 L or 0.5 L vessels. Instructions cover inoculation, medium exchange, sampling, morphology assessment, metabolite and pH analysis, cell counting, viability testing, and harvest in the PBS-Mini bioreactor. The document presents recommended starting parameters for seeding density, working volume, agitation…
This publication investigates hydrodynamic scale-up of human pluripotent stem cell aggregate cultures in 100 mL and 500 mL Vertical-Wheel® Bioreactors. Three-dimensional flow simulations and aggregate trajectory analysis were used to evaluate shear stress and energy dissipation rate exposure across multiple agitation conditions. Exposure profiles were compared with aggregation efficiency, cell growth, aggregate size, and fold expansion during six-day cultures. Energy dissipation rate exposure correlated with aggregate formation and expansion performance, while shear stress was not a…
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